Acute and chronic toxicity of nickel to nitrifiers at different temperatures

This study investigated the acute nickel toxicity on nitrification of low ammonia synthetic wastewater at 10, 23, and 35°C. The nickel inhibition half-velocity constants(K_(I,Ni)) for ammonia oxidizing bacteria(AOB) and nitrite oxidizing bacteria(NOB) based on Ni/MLSS ratio at 10, 23, and 35°C were 5.4 and 5.6 mg Ni/g MLSS, 4.6 and 3.5 mg Ni/g MLSS, and 9.1 and 2.7 mg Ni/g MLSS, respectively. In addition, chronic toxicity of nickel to nitrification of low ammonia synthetic wastewater was investigated at 10°C in two sequencing batch reactors(SBRs). Long-term SBRs operation and short-term batch tests were comparable with respect to the extent of inhibition and corresponding Ni/MLSS ratio. The μ_(max), b, and K_o of AOB were 0.16 day^(-1), 0.098 day^(-1) and 2.08 mg O_2/L after long-term acclimatization to nickel of 1 mg/L at 10°C, high dissolved oxygen(DO)(7 mg/L) and long solids retention time(SRT) of 63–70 days. Acute nickel toxicity of nitrifying bacteria was completely reversible.

Effects of long-term elevated CO_2 on N_2-fixing,denitrifying and nitrifying enzyme activities in forest soils under Pinus sylvestriformis in Changbai Mountain

A study was conducted to determine the effects of elevated CO2 on soil N process at Changbai Mountain in Jilin Province, northeastern China （42°24＂N, 128°06＂E, and 738 m elevation）. A randomized complete block design of ambient and elevated CO2 was established in an open-top chamber facility in the spring of 1999. Changpai Scotch pine （Pinus sylvestris var. sylvestriformis seeds were sowed in May, 1999 and CO2 fumigation treatments began after seeds germination. In each year, the exposure started at the end of April and stopped at the end of October. Soil samples were collected in June and August 2006 and in June 2007, and soil nitrifying, denitrifying and N2-fixing enzyme activities were measured. Results show that soil nitrifying enzyme activities （NEA） in the 5-10 cm soil layer were significantly increased at elevated CO2 by 30.3% in June 2006, by 30.9% in August 2006 and by 11.3% in June 2007. Soil denitrifying enzyme activities （DEA） were significantly decreased by elevated CO2 treatment in June 2006 （P 〈 0.012） and August 2006 （P 〈 0.005） samplings in our study; no significant difference was detected in June 2007, and no significant changes in N2-fixing enzyme activity were found. This study suggests that elevated CO2 can alter soil nitrifying enzyme and denitrifying enzyme activities.

Effect of nitrification inhibitor DMPP on nitrogen leaching, nitrifying organisms, and enzyme activities in a rice-oilseed rape cropping system

DMPP （3,4-dimethylpyrazole phosphate） has been used to reduce nitrogen （N） loss from leaching or denitrification and to improve N supply in agricultural land. However, its impact on soil nitrifying organisms and enzyme activities involved in N cycling is largely unknown. Therefore, an on-farm experiment, for two years, has been conducted, to elucidate the effects of DMPP on mineral N （NH4^＋- N and NO3^--N） leaching, nitrifying organisms, and denitrifying enzymes in a rice-oilseed rape cropping system. Three treatments including urea alone （UA）, urea ＋ 1% DMPP （DP）, and no fertilizer （CK）, have been carded out. The results showed that DP enhanced the mean NH4^＋-N concentrations by 19.1%-24.3%, but reduced the mean NO3^--N concentrations by 44.9%-56.6% in the leachate, under a two-year rice-rape rotation, compared to the UA treatment. The population of ammonia oxidizing bacteria, the activity of nitrate reductase, and nitrite reductase in the DP treatment decreased about 24.5%-30.9%, 14.9%-43.5%, and 14.7%-31.6%, respectively, as compared to the UA treatment. However, nitrite oxidizing bacteria and hydroxylamine reductase remained almost unaffected by DMPP. It is proposed that DMPP has the potential to either reduce NO3^--N leaching by inhibiting ammonia oxidization or N losses from denitrification, which is in favor of the N conversations in the rice-oilseed rape cropping system.

Start-up of anaerobic ammonia oxidation bioreactor with nitrifying activated sludge

The anaerobic ammonia oxidation(Anammox) bioreactor was successfully started up with the nitrifying activated sludge. After anaerobically operated for 105 d, the bioreactor reached a good performance with removal percentage of both ammonia and nitrite higher than 95% and volumetric total nitrogen removal as high as 149.55 mmol/(L·d). The soft padding made an important contribution to the high efficiency and stability because it held a large amount of biomass in the bioreactor.

Performance of Anammox granular sludge bed reactor started up with nitrifying granular sludge

The anaerobic ammonia oxidation(Anammox) granular sludge bed reactor was started up successfully with nitrifying granular sludge. During the operation, the nitrifying granular sludge was gradually converted into Anammox granular sludge with good settling property and high conversion activity. The Anammox reactor worked well with the shortest HRT of 2 43 h. Under the condition that HRT w as 6 39 h and influent concentration of ammonia and nitrite was 10 mmol/L, the removal of ammonia and nitrite was 97 17% and 100 00%, respectively. Corresponding volumetric total nitrogen loading rate and volumetric total nitrogen conversion rate were 100 83 mmol/(L·d) and 98 95 mmol/(L·d). The performance of Anammox reactor was efficient and stable.

Enhanced biological nutrient removal by the alliance of a heterotrophic nitrifying strain with a nitrogen removing ecosystem

Nitrogen removal from synthetic wastewater was investigated in an airlift bioreactor （ALB）, augmented with a novel heterotrophic nitrifier Pseudonocardia ammonioxydans H9^T under organic carbon to nitrogen ratios （Corg/N） ranging from 0 to 12. Effect of the inoculated strain was also determined on the settling properties and the removal of chemical oxygen demand （COD）. Two laboratory scale reactors were set up to achieve a stable nitrifying state under the same physicochemical conditions of hydraulic retention time （HRT）, temperature, pH and dissolved oxygen （DO）, and operated under the sequencing batch mode. The level of DO was kept at 0.5- 1.5 mg/L by periodic stirring and aeration. Each specific Corg/N ratio was continued for duration of 3 weeks. One of the reactors （BR2） was inoculated with P ammonioxydans H9^T periodically at the start of each Corg/N ratio. Sludge volumetric index （SVI） improved with the increasing Corg/N ratio, but no significant difference was detected between the two reactors. BR2 showed higher levels of nitrogen removal with the increasing heterotrophic conditions, and the ammonia removal reached to the level of 82%-88%, up to10% higher than that in the control reactor （BR1） at Corg/N ratios higher than 6; however, the ammonia removal level in experimental reactor was up to 8% lower than that in control reactor at Corg/N ratios lower than 2. The COD removal efficiency progressively increased with the increasing Corg/N ratios in both of the reactors. The COD removal percentage up to peak values of 88%-94% in BR2, up to 11% higher than that in BR1 at Corg/N ratio higher than 4. The peak values of ammonia and COD removal almost coincided with the highest number （18%-27% to total bacterial number） of the exogenous bacterium in the BR2, detected as colony forming units （CFU）. Furthermore, the removal of ammonia and COD in BR2 was closely related to the number of the inoculated strain with a coefficient index （R2） up to 0.82 and 0.85 for ammonia and COD, respectively. These results suggest that it was more efficient for both the ammonia and carbon nutrient removals in a reactor inoculated with a heterotrophic nitrifier at high Corg/N ratio, inferring that the heterotrophic nitrifers would be practically more available in the treatment of wastewater with high level of ammonia and COD.

The Short-term Effects of Temperature and Free Ammonia on Ammonium Oxidization in Granular and Floccular Nitrifying System

The short-term effects of temperature and free ammonia (FA) on ammonium oxidization were investigated in this study by operating several batch tests with two different partial nitrification aggregates, formed as either granules or flocs. The results showed that the rate of ammonium oxidation in both cultures increased significantly as temperature increased from 10 to 30 °C. The specific ammonium oxidation rate with the granules was 2-3 times higher than that with flocs at the same temperature. Nitrification at various FA concentrations and temperatures combination exhibited obvious inhibition in ammonium oxidation rate when FA was 90 mg·L 1 and tempera- ture dropped to 10 °C in the two systems. However, the increase in substrate oxidation rate of ammonia at 30 °C was observed. The results suggested that higher reaction temperature was helpful to reduce the toxicity of FA. Granules appeared to be more tolerant to FA attributed to the much fraction of ammonia oxidizing bacteria (AOB) and higher resistance to the transfer of ammonia into the bacterial aggregates, whereas in the floc system, the bacteria distributed throughout the entire aggregate. These results may contribute to the applicability of the nitrifying granules in wastewater treatment operated at high ammonium concentration.

Purification and Characterization of a Low-temperature Hydroxylamine Oxidase from Heterotrophic Nitrifier Acinetobacter sp.Y16

Objective To purify a low-temperature hydroxylamine oxidase （HAO） from a heterotrophic nitrifying bacterium Acinetobacter sp. Y26 and investigate the enzyme property. Methods A HAO was purified by an anion-exchange and gel-filtration chromatography from strain Y16. The purity and molecular mass were determined by RP-HPLC and SDS-PAGE. The HAO activity was detected by monitoring the reduction of potassium ferricyanide using hydroxylamine as substrate and ferricyanide as electron acceptor. The partial amino acid sequence was determined by mass spectrometry. Results The low-temperature HAO with a molecular mass of 61 kDa was purified from strain Y26 by an anion-exchange and gel-filtration chromatography. The enzyme exhibited an ability to oxidize hydroxylamine in wide temperature range （4-40 ℃） in vitro using hydroxylamine as substrate and ferricyanide as electron acceptor. It was stable in the temperature range of 4 to 25 ℃ and pH range of 6.0 to 8.5 with less than 30% change in its activity. The optimal temperature and pH were 15 ℃ and 7.5, respectively. Three peptides were determined by mass spectrometry which were shown to be not identical to other reported HAOs. Conclusion This is the first study to purify a low-temperature HAO from a heterotrophic nitrifier Acinetobecter sp. It differs from other reported HAOs in molecular mass and enzyme properties. The findings of the present study have suggested that the strain Y26 passes through a hydroxylamine-oxidizing process catalyzed by a low-temperature HAO for ammonium removal.

Effect of organic carbon on nitrification effciency and community composition of nitrifying biofilms

The effects of organic carbon/inorganic nitrogen （C/N） ratio on the nitrification processes and the community shifts of nitrifying biofilms were investigated by kinetic comparison and denaturing gradient gel electrophoresis （DGGE） analysis. The results showed that the nitrification rate decreased with an increasing organic concentration. However, the effect became weak when the carbon concentration reached a sufficiently high level. Denitrification was detected after organic carbon was added. The 12 h ammonium removal rate ranged from 85% to 30% at C/N = 0.5, 1, 2, 4, 8, and 16, as compared to the control （C/N = 0）. The loss of nitrogen after 24 h at C/N = 0.5, 1, 2, 4, 8, and 16 was 31%, 18%, 24%, 65%, 59%, and 62%, respectively. The sequence analysis of 16S rRNA gene fragments revealed that the dominant populations changed from nitrifying bacteria （Nitrosomonas europaea and Nitrobacter sp.） to denitrifying bacteria （Pseudomonas sp., Acidovorax sp. and Comamonas sp.） with an increasing C/N ratio. Although at high C/N ratio the denitrifying bacteria were the dominant populations, nitrifying bacteria grew simultaneously. Consequently, nitrification process coexisted with denitrification.

A Depth Wise Diversity of Free Living N₂Fixing and Nitrifying Bacteria and Its Seasonal Variation with Nitrogen Containing Nutrients in the Mangrove Sediments of Sundarban, WB, India

Mangrove provides a unique ecological niche to different microbes which play various roles in nutrient recycling as well as various environmental activities. The highly productive and diverse microbial community living in mangrove ecosystems continuously transforms dead vegetation and recycle nitrogen, phosphorus, sulphur and other nutrients that can later be used by the plants. Mangrove ecosystems are rich in organic matter, and however, in general, they are nutrient-deficient ecosystems, especially of nitrogen and phosphorus. The present study investigated depth wise variation of Nitrifying bacteria, Nitrogen fixing bacteria, total bacterial population along with nitrate-nitrogen, nitrite-nitrogen and other physicochemical parameters of soil during pre-monsoon, monsoon and post-monsoon periods at three different sampling stations of mangrove sediments viz. deep forest region, rooted region and unrooted region. The microbial population was also found maximum in the deep forest sediment relative to the other two sites. Populations of cultureable microbes were found maximum in surface soil and decreased with increase in depth in Sundarban mangrove environment. A decreasing trend of total microbial load, nitrifying and nitrogen fixing bacteria with increase in depth were recorded throughout the year. Present study revealed the relationship among depth integrated variations of physicochemical components (viz. soil temperature, pH, salinity, nitrite nitrogen and nitrate nitrogen concentration) and total microbial load, nitrifying and nitrogen fixing bacteria microbial populations.

Isolation and Characteristics of New Heterotrophic Nitrifying Bacteria

The study presented the method for isolating the heterotrophic nitrifiers and the characterization of heterotrophic nitrification. When influent Ammonia nitrogen concentration was 42. 78 - 73. 62 mg/L. The average ammonia nitrogen removal rate was 81.32% from the bioceramics reactor. Sodium acetate and ammonium chloride were used as carbon and nitrogen source. The COD removal rates by microorganisms of strain wgy21 and wgy36 were 56.1% and 45.45%, respectively. The TN removal rates by microorganisms of strain wgy21 and wgy36 were 65.85% and 67. 98%, respectively. At the same time, the concentration of ammonium nitrogen was with the removal rates of 75.25% and 84.96%, and it also had the function of producing NO2 - N. Sodium acetate and sodium nitrite were used as carbon and nitrogen source. Through the 12 days of the aerobic culture, the COD femoral rates by microorganisms of strain wgy21 and wgy36 were 29.25% and 22.08%, respectively. NO2 -N concentration decreased slowly. Comparison, similarity of wgy21 and many Acinetobacter sp. ≥99%, similarity of wgy36 and many Acinetobacter sp. ≥99%. Refer to routine physiologicalbiochemical characteristic determination, further evidences showed that wgy21 and wgy36 belong to Acinetobacter sp., respectively.

Biomass Fraction of Phosphate-Accumulating Organisms Grown in Anoxic and Aerobic Stages under Optimum Nitrate Concentration

The effects of nitrate concentration on the capability of phosphorus uptake in the main anoxic stage were investigated.Meanwhile, the biomass fractions — heterotrophs, phosphateaccumulating organisms( PAOs),and nitrifying organisms in a pilot-scale enhanced biological phosphorus removal( EBPR) system— were both experimentally and theoretically evaluated( from the mass balance calculations of organic matter, nitrogen and phosphorus),under optimum nitrate concentration in the main anoxic stage,in which the influent chemical oxygen demand( COD)concentration was stabilized at( 290 ± 10) mg·L- 1and the influent total phosphorus( TP) concentration was stabilized at( 7. 0 ± 0. 5)mg · L- 1. In long term operations,the process exhibited high performance in removing organic matter, nitrogen, and phosphorus. Approximately 46. 41% of organic matter,57. 21% of nitrogen,and 48. 14% of phosphorus were removed from the influent in the form of carbon dioxide,nitrogen gas,and polyphosphate,respectively. XH( heterotrophs),XPAO( PAOs),and XAUT( autotrophs) were regarded as the major organisms responsible for biomass production. The yield fractions of XHgrowth in the first anoxic,the second anoxic,and the aerobic stages were 10. 24%,19. 11%,and 19. 71%,respectively; the yield fractions of XPAO growth in the second anoxic and the aerobic stages were 24. 34% and19. 86%,respectively; the yield fraction of XAUTgrowth in the aerobic stage was 6. 74%. These results showed that XHand XPAOformed the major community. Moreover,a higher amount of XPAOgrowth on stored poly-hydroxyalkanoates( PHAs) under the anoxic condition was seen in this EBPR system for municipal wastewater treatment.

Performance Evaluation of Collard Based Bio-Filter for Treating Fish Tank Effluent for Re-use

The increasing Uganda’s urban population growth has led to limited space coupled with high cost of living, thus making it difficult for the urban poor in congested areas to afford fish protein hence poor nutrition among the low income earners. Therefore this study was conducted to evaluate the performance of collard based bio-filtration system for filtering fish tank effluent for re-use in congested peri-urban areas. Field physical-chemical parameters (ammonia, nitrate, dissolved oxygen, temperature and pH) were measured at various bio-filter lengths in the effluent from both collard based and GBF (Gravel Based Bio-Filter). Differences in mean ammonia and nitrate levels at various lengths were analyzed using one-way ANOVA at p = 0.05. Ammonia levels were significantly reduced (p < 0.05) at various lengths: L0 99.1 mg/L;L1 75.8 mg/L (23.6%);L2 53.1 mg/L (46.4%);L3 25.8 mg/L (74%) and L4 6.6 mg/L (93.4%). Similarly, nitrate levels significantly reduced (p < 0.05): L0 11.8 mg/L;L1 7.2 mg/L (39.4%);L2 3.6 mg/L (69.2%);L3 1.6 mg/L (86.7%) and L4 0.1 mg/L (99.3%). Significant difference (p < 0.05) was obtained in mean ammonia and nitrate removal between collard based and gravel bio-filters. Collard based filter yielded higher ammonia and nitrate removal at L4 by 18.3% and 39.5% respectively, hence L4 is the effective length for collard based bio-filter.

Preservation Property and Decay Kinetic of Polyurethane Immobilized Nitrifying Bacteria Pellets

The preservation methods of polyurethane immobilized nitrifying bacteria pellets which had been enriched in laboratory were provided. Factors such as temperature, pH and light, which affect the nitrification activity of polyurethane immobilized pellets, were investigated. The result showed that dark, deionized water and low temperature is suitable for polyurethane immobilized nitrifying bacteria pellets’ long term preservation.

Removal Effect of Nitrifying Bacteria on Ammonia Nitrogen in Water

[Objectives]To investigate the removal effect of nitrifying bacteria on ammonia nitrogen in water.[Methods]In this experiment,the treated water(referred to as raw water hereinafter)from the Changping Town Qiaoli Water Treatment Project in Dongguan City of Guangdong Province was used as the experimental water body,and the nitrifying bacteria liquid used in the project was taken as the experimental material,to explore the removal effect of the nitrifying bacteria liquid on the ammonia nitrogen in the water body.[Results]Under the condition that other variables remain unchanged,the more the amount of nitrifying bacteria liquid added,the higher the removal efficiency of nitrifying bacteria liquid;under the same conditions,the removal effect of ammonia nitrogen in a stirred water body is better than that in an unstirred water body;the removal effect of ammonia nitrogen in a water body with bio-media/carriers is better than that without bio-media/carriers.[Conclusions]Nitrifying bacteria have a better removal effect on the ammonia nitrogen in the water body.

Impact of 17β-Estradiol on Natural Water’s Heterotrophic Nitrifying Bacteria

In this research, bottom water samples were collected from nature water. After cultivating and selecting, bacteria which could use (NH4)2SO4 as the only nitrogen source had been selected. The bacteria were cultivated in BM cultures with 0, 0.1, 1, 10, 100 ng/L 17β-estradiol (E2), and the initial concentration of E2 is the only difference between cultures of each group. BM culture is a kind of bacteria culture with 100 mg/L of NH4-N as only nitrogen source. Every group’s N- NH4+, N- NO3-concentration and OD600 were measured. The result shows that compared with the control group, in which no E2 was added, the growth of heterotrophic nitrifying bacteria had been promoted when the concentration of E2 was in range of 1 - 100 ng/L. In addition, heterotrophic nitrifying bacteria’s growing speed has a positive correlation between the E2’s concentration. However, low concentration of E2 (like 0.1 ng/L), could inhibit the growth of heterotrophic nitrifying bacteria. Considering the impact of E2 on heterotrophic nitrifying bacteria, it is necessary to intensify the detection of E2 in the future.

The response of structure and nitrogen removal function of the biofilm on submerged macrophytes to high ammonium in constructed wetlands

The ammonium exceedance discharge from sewage treatment plants has a great risk to the stable operation of subsequent constructed wetlands(CWs).The effects of high ammonium shocks on submerged macrophytes and epiphytic biofilms on the leaves of submerged macrophytes in CWs were rarely mentioned in previous studies.In this paper,the 16S rRNA sequencing method was used to investigate the variation of the microbial communities in biofilms on the leaves of Vallisneria natans plants while the growth characteristics of V.natans plants were measured at different initial ammonium concentrations.The results demonstrated that the total chlorophyll and soluble sugar synthesis of V.natans plants decreased by 51.45%and 57.16%,respectively,and malondialdehyde content increased threefold after8 days if the initial NH_(4)^(+)-N concentration was more than 5 mg/L.Algal density,bacterial quantity,dissolved oxygen,and pH increased with high ammonium shocks.The average removal efficiencies of total nitrogen and NH_(4)^(+)-N reached 73.26%and 83.94%,respectively.The heat map and relative abundance analysis represented that the relative abundances of phyla Proteobacteria,Cyanobacteria,and Bacteroidetes increased.The numbers of autotrophic nitrifiers and heterotrophic nitrification aerobic denitrification(HNAD)bacteria expanded in biofilms.In particular,HNAD bacteria of Flavobacterium,Hydrogenophaga,Acidovorax,Acinetobacter,Pseudomonas,Aeromonas,and Azospira had higher abundances than autotrophic nitrifiers because there were organic matters secreted from declining leaves of V.natans plants.The analysis of the nitrogen metabolic pathway showed aerobic denitrification was the main nitrogen removal pathway.Thus,the nitrification and denitrification bacterial communities increased in epiphytic biofilms on submerged macrophytes in constructed wetlands while submerged macrophytes declined under ammonium shock loading.

The short and long-term effect of polystyrene nanoplastics on nitrifying sludge at high nitrite concentrations

The effect of nanoplastics(NPs)on nitrite oxidation bacteria(NOB)community in treating high-strength wastewater remains unclear,which seriously affects the stability of nitrogen removal process.In this study,highly active nitrifying sludge was enriched and exposed to 50nm polystyrene NPs(PS-NPs)for short-term(1,100,500,and 1000 mg/L,1.5 hr)and long-term(1,10,100 mg/L,40 days)at high nitrite concentration.In contrast to previous studies,our results showed that the exposures to PS-NPs had little effect on nitrifying performances.After long-term exposure,the protein/polysaccharide ratios in extracellular polymeric substances(EPS)were positively correlated with PS-NPs concentrations(0.78–0.99).The produced reactive oxygen species(ROS)were gradually removed,and PS-NPs higher than 10 mg/L caused damage to membrane integrity.Long-term exposure for 40 days increased the community diversity and caused significant differences between the control and exposed communities.The control group were dominated by Nitrobacter and Exiguobacterium,while the exposure group was dominated by Bacillus,Mycobacterium,and Nitrospira.A noticeable shift in the NOB community from Nitrobacter(26.5%to 3.4%)to Nitrospira(1.61%to 14.27%)was observed.A KEGG analysis indicated a decrease in cell growth and death,cell motility and energy metabolism.It appeared that NOB could adapt to PS-NPs stress through enhanced secretion and removal of oxidative damage.Overall,this study provided new insights into the response mechanism of NOB to PS-NPs exposure.

Insights into effects of thermotolerant nitrifying and sulfur-oxidizing inoculants on nitrogen-sulfur co-metabolism in sewage sludge composting

In this study,high temperature thermotolerant nitrifying bacteria(TNB)and high temperature thermotolerant sulfide oxidizing bacteria(TSOB)were obtained from compost samples and inoculated into sewage sludge(SS)compost.The effects of inoculation on physical and chemical parameters,ammonia and hydrogen sulfide release,nitrogen form and sulfur compound content change and physical-chemical properties during nitrogen and sulfur conversion were studied.The results showed that inoculation of TNB and TSOB increased the temperature,pH,OM degradation,C/N ratio and germination index(GI)of compost.Compared with the control treatment(CK),the addition of inoculants reduced the release of NH3 and H2S,and transformed them into nitrogen and sulfur compounds,the hydrolysis of polymeric ferrous sulfate was promoted,resulting in relatively high content of sulfite and sulfate.At the same time,the physical and chemical properties of SS have a strong correlation with nitrogen and sulfur compounds.

Differences in distributions, assembly mechanisms, and putative interactions of AOB and NOB at a large spatial scale

Ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) play crucial roles in removing nitrogen from sewage in wastewater treatment plants (WWTPs) to protect water resources. However, the differences in ecological properties and putative interactions of AOB and NOB in WWTPs at a large spatial scale remain unclear. Hence, 132 activated sludge (AS) samples collected from 11 cities across China were studied by utilizing 16S rRNA gene sequencing technology. Results indicated that Nitrosomonas and Nitrosospira accounted for similar ratios of the AOB community and might play nearly equal roles in ammonia oxidation in AS. However, Nitrospira greatly outnumbered other NOB genera, with proportions varying from 94.7% to 99.9% of the NOB community in all WWTPs. Similar compositions and, hence, a low distance–decay turnover rate of NOB (0.035) across China were observed. This scenario might have partly resulted from the high proportions of homogenizing dispersal (~13%). Additionally, drift presented dominant roles in AOB and NOB assembling mechanisms (85.2% and 81.6% for AOB and NOB, respectively). The partial Mantel test illustrated that sludge retention time and temperature were the primary environmental factors affecting AOB and NOB communities. Network results showed that NOB played a leading role in maintaining module structures and node connections in AS. Moreover, most links between NOB and other microorganisms were positive, indicating that NOB were involved in complex symbioses with bacteria in AS.